Michel Abely

Broad respiratory virus detection in infants hospitalized for bronchiolitis by use of a multiplex RT-PCR DNA microarray system†

Newly available molecular tools allow a sensitive detection of a broad panel of viruses in respiratory tract specimens. In the present study, the application of a multiplex RT‐PCR DNA microarray in diagnosis and epidemiological survey of viral infections in infants hospitalized for bronchiolitis was assessed. One hundred and thirty‐eight nasopharyngeal aspirates collected from October 2007 to September 2008 were tested by direct immunofluorescence and viral culture, a combination of referenced RT‐PCRs and the DNA microarray. One or more viruses were detected in 96, 126 and 126 of the specimens by direct immunofluorescence and viral culture, RT‐PCRs and DNA microarray, respectively (70 vs. 91 vs. 91%, P < 10−3). The RT‐PCRs and the DNA microarray yielded concordant results for 99% of specimens and identified mixed viral infections in 85 (62%). The most common associations were: human bocavirus and respiratory syncytial virus (32%), adenovirus and respiratory syncytial virus (30%), and parainfluenza virus type 3 and respiratory syncytial virus (23%). None of the bronchiolitis severity parameters including intensive care unit admission, O2 supply, O2 saturation percentage, O2 length and length of stay at the hospital appeared to be significantly increased in multiple viral infections compared to single viral infections (P > 0.1). In conclusion, the use of this DNA microarray in clinical virology practice allows rapid and accurate identification of common and uncommon viral respiratory pathogens in infants hospitalized for bronchiolitis. It should improve the clinical management, the epidemiological survey, and the prevention of the nosocomial transmission of respiratory viruses in pediatric wards. J. Med. Virol. 84:979–985, 2012.

Metabolic control in children with diabetes mellitus who are younger than 6 years at diagnosis: continuous subcutaneous insulin infusion as a first line treatment?

OBJECTIVE To assess long-term metabolic outcomes in children with diabetes mellitus that was diagnosed when they were <6 years old. STUDY DESIGN A cohort of 66 children with diabetes mellitus that had a duration of at least 5 years and was diagnosed before they were 6 years old. Thirty-four children were treated at diagnosis with multiple daily subcutaneous insulin injections (MDI), and all these children, except 3, were switched to continuous subcutaneous insulin infusion (CSII; group A). Thirty-two children received CSII as initial treatment (group B). RESULTS Hemoglobin A1c values were significantly lower in patients receiving CSII than MDI during all the 8 years of follow-up except one (year 1: 6.9%+/-0.9% versus 7.6%+/-1%, P=.011 ; year 4: 7.4%+/-0.8% versus 8.1%+/-0.9%, P=.006; year 7: 7.6%+/-0.5% versus 8.3%+/-0.8%, P=.001). The incidence of severe hypoglycemia was greatly decreased for the CSII group (9.8 versus 22.3 episodes/100 patient-years, P=.016). In group A, hemoglobin A1c values increased during the study period, and in group B, they increased only during the first 2 years and remained constant thereafter. Only 9.1% of patients did not use or abandoned CSII. CONCLUSION CSII in children<6 years of age enables better long-term metabolic control and lowers the risk of severe hypoglycemia better than MDI, especially when initiated at diagnosis.

Causes of death in French cystic fibrosis patients: The need for improvement in transplantation referral strategies!

BACKGROUND Little data exist on causes of death in cystic fibrosis (CF) patients in the era of lung transplantation. METHODS Deaths in CF patients in France (2007-2010) were identified using the French CF Registry and causes of deaths were determined based on medical files by a mortality adjudication committee. RESULTS Of 256 deaths, half occurred after lung transplantation and were related to early or late complications of transplantation, whereas half occurred in patients who did not receive lung transplantation and were primarily related to respiratory failure or massive hemoptysis. Among patients who did not receive lung transplantation, only 19% died while waiting on a lung transplantation list. Lack of listing for lung transplantation was primarily related to late, or to lack of transplantation referral, rather than to contraindication to transplantation. CONCLUSIONS These data suggest that improvement in transplantation referral strategies may result in transplantation-related survival benefits.

Cystic fibrosis airway epithelium remodelling: involvement of inflammation

Chronic inflammation is a hallmark of cystic fibrosis (CF) lung disease and airway epithelium damage and remodelling are important components of lung pathology progression in CF. Whether this remodelling is secondary to deleterious infectious and inflammatory mediators, or to alterations of CF human airway epithelial (HAE) cells, such as their hyper inflammatory phenotype or their basic cystic fibrosis transmembrane conductance regulator (CFTR) default, remains debated. In this study, we evaluated the involvement of alterations of CF HAE cells in airway epithelium remodelling. HAE cells from non‐CF and CF patients were cultured in an air–liquid interface, with and without inflammatory stimulation, along the regeneration process, and the remodelling of the reconstituted epithelium was analysed. We confirmed that CF HAE cells showed a hyperinflammatory phenotype which was lost with time. In comparison to non‐CF epithelium, CF epithelium regeneration in the absence of exogenous inflammation was higher and exhibited basal cell hyperplasia. This remodelling was mimicked by inflammatory stimulation of non‐CF cells and was absent when CF HAE cells were no longer hyperinflamed. Moreover, the number of goblet cells was similar in non‐CF and CF cultures and increased equally under inflammatory stimulation. Finally, whatever the inflammatory environment, CF cultures showed a delay in ciliated cell differentiation. In conclusion, alterations of CF HAE cells partly regulate airway epithelium remodelling following injury and regeneration. This remodelling, together with goblet cell hyperplasia induced by exogenous inflammation and alteration of ciliated cell differentiation, may worsen mucociliary clearance impairment, leading to injury. Copyright

Phylogenetic analysis of Echovirus 30 isolated during the 2005 outbreak in France reveals existence of multiple lineages and suggests frequent recombination events

BACKGROUND Echovirus 30 (E-30) was responsible in France for a major aseptic meningitis outbreak during 2005 summer season. However, the virological mechanisms responsible for the periodic emergence of the epidemic strains remain to be investigated. OBJECTIVES To assess the genetic diversity of two genome regions, VP1 and 3Dpol, of echovirus 30 strains isolated during the 2005 aseptic meningitis outbreak in Champagne Ardenne (CA) area (France). STUDY DESIGN Partial VP1 genomic region of 23 E-30 strains isolated in CA was sequenced and compared with 73 E-30 strains originating from different French areas to estimate the number and the diversity of E-30 lineages. Partial sequences for 3D polymerase (3Dpol) were analyzed to detect potential recombination events within the non-structural (NS) region of the genome of EV neurotropic strains. RESULTS Phylogenetic analysis of the VP1 evidenced the co-circulation of 6 distinct E-30 lineages responsible for the 2005 aseptic meningitis outbreak in France of which three had co-circulated in CA. Partial sequencing of the 3Dpol coding region showed that all of the E-30 strains exhibited different phylogenetic links between VP1 and 3Dpol genomic regions, suggesting multiple intra- or inter-serotypic recombination events within the NS part of the genome. CONCLUSIONS Our findings revealed existence of multiple lineages and suggested frequent recombination events among E-30 strains having co-circulated in a restricted area during a short time outbreak period. Moreover, our data demonstrated that study of single VP1 genome region analysis could not accurately describe the phylogenetic origin of E-30 isolates.

Rhinovirus‐associated pulmonary exacerbations show a lack of FEV1 improvement in children with cystic fibrosis

Respiratory viral infections lead to bronchial inflammation in patients with cystic fibrosis, especially during pulmonary exacerbations. The aim of this study was to determine the impact of viral‐associated pulmonary exacerbations in children with cystic fibrosis and failure to improve forced expiratory volume in 1 s (FEV1) after an appropriate treatment.

Four family cases of acute renal failure: question

An 11 year-old boy was referred and admitted to our institution for acute kidney injury (AKI). He had no significant past medical events and no family history of kidney disease. Before his admission, he had experienced vomiting and diarrhea for 4 days, leading to oliguria. Because these symptoms were shared with his parents and grandparents, acute viral gastroenteritis was clinically suspected and symptomatic treatment was started. Despite this symptomatic treatment, vomiting and diarrhea persisted. The child was referred to the pediatric emergency department of our hospital. At the time of admission, his physical examination was normal. He had no fever or edema, and his blood pressure was 142/ 72 mmHg. Urinary tests showed microscopic hematuria and proteinuria <1 g/L. His laboratory results were as follows: serum creatinine 12.9 mg/dl (1,137 μmol/L); sodium (Na) 123 mmol/L; potassium (K) 9.1 mmol/L; bicarbonate (HCO3 ) 13 mmol/L. His electrocardiogram

Moyamoya disease associated with hereditary spherocytosis.

A 5-year-old girl with hereditary spherocytosis presented with two episodes of transient ischemic attacks within a month. Cranial magnetic resonance imaging angiography revealed a left internal carotid artery and middle cerebral artery stenosis, with an extensive vascular mesh in the thalamic area indicative of moyamoya disease. Treatment consisted of supporting cerebral perfusion with blood transfusions, and splenectomy to prevent recurrence. Moyamoya disease is a very unusual cerebrovascular disorder in childhood and its association with hereditary spherocytosis is rarely reported.

WS07.5 Rhinovirus associated pulmonary exacerbations show a lack of FEV1 improvement in children with cystic fibrosis

Objectives Respiratory viral infections lead to bronchial inflammation in cystic fibrosis patients, especially during pulmonary exacerbations. The aim of the present study was to determine the impact of viral associated pulmonary exacerbations in children with cystic fibrosis and failure to improve force expiratory volume in 1 second (FEV1) levels after an appropriate treatment. Methods Children with a diagnosis of cystic fibrosis were longitudinally evaluated three times: at baseline (Visit 1), at the diagnosis of pulmonary exacerbation (Visit 2) and after exacerbation treatment (Visit 3). Nasal and bronchial swabs were analyzed at each visit with multiplex viral respiratory PCR panel. Pulmonary function tests were recorded at each visit, in order to highlight a possible failure to improve them after treatment. Lack of improvement was defined by an increase of FEV1 less than 5% between Visit 2 and Visit 3. Results 18 children were included in the study. 10 patients failed to show an improvement of more than 5% in their FEV1 between Visit 2 and Visit 3. Rhinovirus infection at Visit 2 or Visit 3 was the only risk factor significantly associated with such a failure, both in univariate (OR 12, p Conclusion Rhinovirus infection seems to play a role in the FEV1 recovery after pulmonary exacerbation treatment in children with cystic fibrosis. Such an association needs to be confirmed by a large-scale study, since this finding may have important implication on pulmonary exacerbation management.

Four family cases of acute renal failure: answer

1. In this report, four family members developed AKI within a few days following gastroenteritis. Their renal biopsies showed acute tubulointerstitial nephritis (TIN). A toxic aetiology was therefore suspected [1]. There was no history of nephrotoxic drug exposure. However, all patients ate wild mushrooms during the previous 2 weeks. Taken together, histological findings and analysis of symptoms supported the hypothesis of an orellanus syndrome secondary to ingestion of poisonous mushrooms from the Cortinarius genus [2]. 2. In cases of mushroom poisoning, the mycological identification is often uncertain and is not sufficient to confirm which mushroom was responsible for the toxicity. Therefore, identification of Cortinarius poisoning is classically performed by detecting the fungal toxin called orellanine in the biological fluids or in renal tissues [3, 4]. The toxin has been detected by high-performance liquid chromatography in renal biopsy samples [4, 5]. However, this analytical method is not generally available in clinical practice. In this report, the diagnosis of orellanus syndrome was confirmed 3 weeks after intoxication by the presence of fungal spores of Cortinarius orellanoides identified by light microscopy in the leftover contaminated meals (Fig. 1). 3. Currently, no specific treatment and no antidote is available for Cortinarius poisoning. The efficacy of corticosteroids and antioxidants (vitamin C, Nacetylcysteine) is not established [2]. Moreover, This article refers to the article that can be found at http://dx.doi.org/ 10.1007/s00467-010-1545-7. D. Talmud : J. Motte :M. Abely :C. Pietrement Pediatric Department, American Memorial Hospital, CHU de Reims, 47-51 rue Cognacq Jay, 51092 Reims, France